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WO2017081750A1 - Système de test de cancer et procédé d'évaluation de test de cancer - Google Patents

Système de test de cancer et procédé d'évaluation de test de cancer Download PDF

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Publication number
WO2017081750A1
WO2017081750A1 PCT/JP2015/081611 JP2015081611W WO2017081750A1 WO 2017081750 A1 WO2017081750 A1 WO 2017081750A1 JP 2015081611 W JP2015081611 W JP 2015081611W WO 2017081750 A1 WO2017081750 A1 WO 2017081750A1
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Prior art keywords
nematode
quality
analysis
chemotaxis
test
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PCT/JP2015/081611
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English (en)
Japanese (ja)
Inventor
坂入　実
中村　拓
光一 寺田
菅谷　昌和
範人 久野
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Hitachi Ltd
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Hitachi Ltd
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Priority to JP2017549902A priority Critical patent/JP6522778B2/ja
Priority to PCT/JP2015/081611 priority patent/WO2017081750A1/fr
Publication of WO2017081750A1 publication Critical patent/WO2017081750A1/fr
Anticipated expiration legal-status Critical
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers

Definitions

  • the present invention relates to a technique for a cancer test system and a cancer test evaluation method for performing a cancer test using a nematode.
  • Patent Document 1 describes a cancer detection method using the olfactory sense of nematodes.
  • the present invention has been made in view of such a background, and an object of the present invention is to easily confirm the quality of nematodes used in cancer testing.
  • the present invention provides a test analysis line in which a nematode group dispensed from the same culture unit into a plurality of analysis units is a nematode group for test chemotaxis analysis for a cancer test.
  • the nematode group is divided into a nematode group for quality analysis, which is a nematode group for quality chemotaxis analysis for quality assurance analysis of nematodes, and what chemotaxis behavior the nematode performs Based on the information output from the quality analysis unit and the quality analysis unit that outputs information on the chemotaxis behavior of the quality analysis nematode group with respect to a reference material that is known, information on the quality of the nematode is obtained.
  • the quality analysis unit Based on the information output from the test analysis unit, the quality analysis unit to output, the test analysis unit that outputs information on the chemotaxis behavior of the test nematode group for the specimen provided from the subject, The chemotaxis result information on the attracting behavior and repelling behavior of the nematode on the specimen Form, based on the run of result information, and having a inspection analysis section for outputting a result of the cancer screening in a subject who provided the sample.
  • Other solutions are described as appropriate in the embodiments.
  • the quality of the nematode used for the cancer test can be easily confirmed.
  • FIG. (1) which shows an example of a specific structure of the quality analysis apparatus which concerns on this embodiment.
  • FIG. (2) which shows an example of the specific structure of the quality analysis apparatus which concerns on this embodiment.
  • FIG. (2) which shows an example of the specific structure of the quality analysis apparatus which concerns on this embodiment.
  • FIG. (1) which shows an example of a specific structure of the quality analysis apparatus which concerns on this embodiment.
  • FIG. (2) which shows an example of the specific structure of the quality analysis apparatus which concerns on this embodiment.
  • FIG. 1 is a diagram illustrating a configuration example of a cancer test system according to the present embodiment.
  • the chemotaxis analysis of the nematode on the subject's urine is performed, and the cancer is based on the nematode's attracting and repelling behavior (the chemotaxis behavior). Test positive / negative for.
  • the chemotaxis analysis of nematodes is performed using a standard substance instead of urine such as isoamyl alcohol, and nematodes used for cancer tests based on the attracting and repelling behavior of nematodes are suitable for the test. Analyzes for quality assurance to determine whether or not.
  • the chemotaxis analysis for testing the subject's urine using the urine of the subject is referred to as the test chemotaxis analysis, and using a urine substitute (standard substance) such as isoamyl alcohol.
  • the chemotaxis analysis for determining whether or not the nematode used in the cancer test is suitable for the test is referred to as quality chemotaxis analysis.
  • the reference material is a material for which it is known whether the nematode performs an attracting action or a repelling action (that is, what kind of chemotaxis behavior the nematode performs), and in this embodiment Use isoamyl alcohol, which is known to cause attracting behavior by nematodes.
  • isoamyl alcohol is used as a standard substance in the quality-taxis analysis.
  • the present invention is not limited to this, and any drug that knows whether a nematode performs an attracting action or a repelling action may be used. That's fine.
  • nematode In the quality chemotaxis analysis in the present embodiment, it is determined whether or not the nematode used for the cancer test is appropriate by analyzing the chemotaxis characteristics of the nematode with respect to isoamyl alcohol. Since nematodes are known to perform an attracting action on isoamyl alcohol, the quality of the nematode can be confirmed by analyzing the attracting action of the nematode on isoamyl alcohol. In the present embodiment, “nematode” refers to a “nematode group” unless otherwise specified.
  • the cancer test system Z includes a quality analysis device (quality analysis unit) 1, a quality analysis device (quality analysis unit) 2, a test analysis device (test analysis unit) 3, and a test analysis device (test analysis unit) 4.
  • nematodes are dispensed from one nematode culture dish 5a (culture unit) into five petri dishes 5b to 5f (a plurality of analysis units).
  • five petri dishes 5b to 5f a plurality of analysis units.
  • four petri dishes 5b to 5e nematodes
  • one petri dish 5f (nematode) is introduced into the quality analysis apparatus 1 for quality chemotaxis analysis (quality analysis nematode group).
  • nematodes are dispensed from one nematode culture petri dish 5a to five petri dishes 5b to 5f, and four petri dishes 5b to 5e are used for examination chemotaxis analysis among the five petri dishes 5b to 5f.
  • the petri dishes 5b to 5e for analysis of test runnability are appropriately referred to as test petri dishes 51.
  • the quality of nematodes used for cancer testing is confirmed by separating nematodes cultured under the same conditions for test chemotaxis analysis and quality chemotaxis analysis. can do.
  • the quality analysis device 1 performs a quality runnability analysis and transmits the obtained result to the quality analysis device 2. Based on the information acquired from the quality analysis device 1, the quality analysis device 2 analyzes information related to the quality of the nematode used for the inspection. The test analysis device 3 performs a test run analysis and transmits the obtained result to the test analysis device 4. The test analysis device 4 analyzes information on the attracting behavior and the avoidance behavior based on the information acquired from the test analysis device 3, and determines whether the cancer is positive or negative. The quality analysis device 2 and the test analysis device 4 output the test result notification 900 in which the test results are described and notified to the subject.
  • FIG. 2 is a functional block diagram illustrating a configuration example of the quality analysis apparatus according to the present embodiment.
  • the quality analysis apparatus 1 includes an imaging unit 101, a placement unit 102, and an illumination unit 103.
  • a petri dish 5f (see FIG. 1) into which nematodes to be subjected to quality motility analysis are dispensed is mounted by an inspector.
  • the illumination unit 103 illuminates the petri dish 5f placed on the placement unit 102 from below.
  • the imaging unit 101 photographs (captures) the petri dish 5f (nematode) placed on the placement unit 102 and illuminated from below by the illumination unit 103.
  • the imaging unit 101 captures a still image with a camera, but a moving image may be captured as a video camera. Note that the imaging unit 101 desirably performs imaging by autofocus. A specific configuration example of the quality analysis apparatus 1 will be described later.
  • FIG. 3 is a functional block diagram illustrating a configuration example of the quality analysis apparatus according to the present embodiment.
  • the quality analysis device 2 is, for example, a PC (Personal Computer) or the like, and includes a memory 201, a storage device 202, a CPU (Central Processing Unit) 203, a communication device 204, an input device 205, and an output device 206.
  • the communication device 204 transmits / receives data to / from the quality analysis device 1 or the like.
  • the input device 205 is a keyboard, a mouse, or the like.
  • the output device 206 is a printer or the like, and prints the inspection result notification 900 (FIG. 1).
  • the storage device 202 is an HD (Hard Disk), SSD (Solid State Drive) or the like, and stores culture condition data 22, quality analysis data 24, nematode data 25, and the like.
  • the culture condition data 22, quality analysis data (information about the quality of nematodes) 24, and nematode data 25 will be described later.
  • a program stored in the storage device 202 is loaded into the memory 201, and the loaded program is executed by the CPU 203, so that the counting unit 211, the calculation unit 212, the determination unit 213, and the output processing unit 214 are loaded. It is embodied.
  • the counting unit 211 indicates the number of nematodes that are attracting to the standard substance (isoamyl alcohol), the number of nematodes that are repelling the standard substance, and the neutral behavior. Count the number of nematodes.
  • neutral behavior is behavior of a nematode that does not show any attracting behavior or repelling behavior.
  • the calculation unit 212 calculates the chemotaxis index CI based on the counting result obtained by the counting unit 211.
  • the chemotaxis index will be described later.
  • the determination unit 213 Based on the chemotaxis index CI calculated by the calculation unit 212, the determination unit 213 performs quality determination as to whether or not the nematode used in the cancer test is an appropriate nematode.
  • the output processing unit 214 prints the determination result by the determination unit 213 on the inspection result notification 900. Note that the counting unit 211, the calculating unit 212, the determining unit 213, and the output processing unit 214 perform the same processing as the counting unit 411, the calculating unit 412, the determining unit 413, and the output processing unit 414 in the test analyzer 4 described later.
  • the counting unit 211, the calculation unit 212, the determination unit 213, and the output processing unit 214 in the quality analysis device 2 and the counting unit 411, the calculation unit 412, the determination unit 413, and the output processing unit 414 in the inspection analysis device 4 are combined. These are appropriately referred to as a counting unit 11, a calculation unit 12, a determination unit 13, and an output processing unit 14.
  • FIG. 4 is a functional block diagram illustrating a configuration example of the inspection analysis apparatus according to the present embodiment.
  • the inspection analysis device 3 includes an observation unit 301, a storage unit 302, and a transport unit 303.
  • the observation unit 301 includes an imaging unit 311, a placement unit 312, and an illumination unit 313.
  • the placement unit 312 is placed with an inspection petri dish 51 (see FIG. 1) into which nematodes to be subjected to the inspection runnability analysis are dispensed.
  • the illumination unit 313 illuminates the inspection petri dish 51 placed on the placement unit 312 from below.
  • the imaging unit 311 photographs the inspection petri dish 51 (nematode) placed on the placement unit 312 and illuminated by the illumination unit 313 from below.
  • the imaging unit 311 captures a still image with a camera, but a moving image may be captured as a video camera. Note that the imaging unit 311 desirably performs imaging by autofocus.
  • the storage unit 302 stores the inspection petri dish 51 for which imaging has been completed for the next imaging.
  • the transport unit 303 transports the inspection petri dish 51 from the observation unit 301 to the storage unit 302 or transports the inspection petri dish 51 from the storage unit 302 to the observation unit 301. In this way, the inspection petri dish 51 is moved to the storage unit 302 after one image is taken, and after a predetermined time has passed after the taking, the inspection petri dish 51 is moved from the storage unit 302 to the mounting unit 312. Imaging is performed by the imaging unit 311. When the imaging is finished, the inspection petri dish 51 is again carried to the storage unit 302 by the transport unit 303. In this way, the chemotactic behavior of the nematode is photographed every certain time.
  • test petri dish 51 is stored in the storage unit 302 for a certain period of time (about 30 minutes) immediately after being placed on the placement unit 312, moved to the observation unit 301 after the end of chemotaxis, and the imaging unit 311 Still images may be taken and the runnability index CI may be calculated.
  • the inspection analysis apparatus 3 may include a discarding unit (not shown) that stores the inspection petri dish 51 that has been completed a predetermined number of times and discarded.
  • the conveyance part 303 carries the inspection petri dish 51 to the disposal part.
  • FIG. 5 is a functional block diagram illustrating a configuration example of the test analyzer according to the present embodiment.
  • the test analysis device 42 is, for example, a PC or the like, and includes a memory 401, a storage device 402, a CPU 403, a communication device 404, an input device 405, and an output device 406.
  • the communication device 404 transmits / receives data to / from the inspection analysis device 3 or the like.
  • the input device 405 is a keyboard, a mouse, or the like.
  • the output device 406 is a printer or the like, and prints an inspection result notification 900 (see FIG. 1).
  • the output device 206 of the quality analysis device 2 and the output device 406 of the inspection analysis device 4 may be a single device.
  • the storage device 402 is an HD, an SSD, or the like, and stores examination analysis data 21, culture condition data 22, and chemotaxis condition data 23.
  • the inspection analysis data 21, culture condition data 22, and chemotaxis condition data 23 will be described later.
  • the culture condition data 22 holds data having the same contents as the culture condition data 22 stored in the storage device 202 of the quality analyzer 2.
  • the memory 401 is loaded with a program stored in the storage device 402, and the loaded program is executed by the CPU 403 so that the counting unit 411, the calculation unit 412, the determination unit 413, and the output processing unit 414 are displayed. It is embodied.
  • the counting unit 411 indicates the number of nematodes showing attracting behavior with respect to the sample (subject's urine), the number of nematodes showing repelling behavior with respect to the sample, and neutral behavior. Count the number of nematodes.
  • the neutral behavior is a behavior of a nematode that does not show an attracting behavior or a repelling behavior.
  • the calculation unit 412 calculates the average value (average chemotaxis index) of the chemotaxis index CI and the chemotaxis index CI based on the count result by the counting unit 411.
  • the chemotaxis index CI will be described later.
  • the determining unit 413 determines negative / positive as a result of the cancer test based on the average value (average chemotaxis index) of the chemotaxis index CI calculated by the calculator 412.
  • the average chemotaxis index will be described later.
  • the output processing unit 414 prints the determination result by the determination unit 412 on the inspection result notification 900.
  • the counting unit 211, the calculation unit 212, the determination unit 213, and the output processing unit 214 in the quality analysis device 2, and the counting unit 411, the calculation unit 412, the determination unit 413, and the output processing unit 414 in the inspection analysis device 4 are used.
  • a counting unit 11 a calculation unit 12, a determination unit 13, and an output processing unit 14.
  • quality analysis apparatus 1 and the inspection analysis apparatus 43 may be integrated into one apparatus.
  • FIGS. 6 and 7 are diagrams illustrating an example of a specific configuration of the quality analysis apparatus according to the present embodiment. 6 shows a state before the nematode is set, and FIG. 7 shows a state after the nematode is set. Note that the quality analysis apparatus 1 shown in FIGS. 6 and 7 is an example, and is not limited to this configuration.
  • a petri dish 5f in which a nematode for quality mobility analysis is dispensed is placed on a plate-like placement portion 102.
  • the placement unit 102 slides along the slide portion 111, and the placement unit 102 is placed on the glass portion 112 (FIG. 6) as shown in FIG. Set.
  • the part under which the petri dish 5f is set is comprised with the transparent member.
  • the glass portion 112 is made of a transparent member.
  • An illumination unit 103 (see FIG. 2) is installed under the glass unit 112. As shown in FIG. 7, when the mounting part 102 is set on the glass part 112, the illumination part 103 will light and the petri dish 5f will be illuminated from the downward direction.
  • the lighting unit 103 may be turned on by an inspector operating a lighting switch provided in the quality analysis apparatus 1. Alternatively, for example, a switch or a sensor is provided at the end of the slide unit 111, and when the mounting unit 102 is set on the glass unit 112, the illumination unit 103 is turned on in response thereto. Good.
  • the mounting portion 102 is joined to the base portion 113, and the annular first support portion 114 is joined to the base portion 113. And the rod-shaped 2nd support part 115 is supported by the form inserted in the annular part of the 1st support part 114.
  • the sandwiching portion 116 is provided around the end of the second support 115 that is different from the end provided with the first support 114.
  • the sandwiching portion 116 is installed on the second support portion 115 by a tightening force by the screw 121.
  • the sandwiching portion 116 is provided with an L-shaped third support portion 117 by, for example, a screw (not shown) or the like, and the third support portion 117 is different from the joint portion with the second support portion 115.
  • An imaging unit 101 is provided at the end.
  • the imaging unit 101 is provided toward the glass unit 112.
  • the petri dish 5f nematode
  • Photographing may be performed by an inspector operating a switch provided in the quality analysis apparatus 1 or may be performed automatically when the illumination unit 103 is turned on.
  • the clamping part 116 becomes movable along the 2nd support part 115 by loosening the screw 121 in the clamping part 116. In this way, the height of the imaging unit 101 can be adjusted.
  • the inspector places the petri dish 5f on the placing part 102 in the state shown in FIG. 6 and sets the placing part 102 on the glass part 112 as shown in FIG. Shoot the nematode). Thereafter, the inspector pulls out the placement unit 102 to return to the state of FIG. 6 and then takes out the petri dish 5f. Then, the inspector places another petri dish 5f on the placement unit 102 and performs imaging in the same procedure. The inspector places the petri dish 5f after a predetermined time on the placement unit 102 again after photographing and performs photographing in the same procedure. By repeating this, an image of a nematode photographed every certain time can be obtained.
  • FIG. 8 is a diagram illustrating an example of inspection analysis data according to the present embodiment.
  • the inspection analysis data 21 includes columns for a lot number, a dispensing number, an inspection mobility result, a culture condition ID (Identification), a mobility condition ID, a quality assurance ID, and an inspection result.
  • the lot number is an identification number uniquely assigned to the nematode culture dish 5a in which nematodes are cultured.
  • the dispensing number is an identification number uniquely given to the dishes 5b to 5f dispensed from the nematode culture dish 5a having the lot number.
  • the petri dish 5b with the dispensing numbers “A1-01”, “A1-02”, “A1-03”, “A1-04”, “A1-05” is started from the petri dish with the lot number “A1”. ⁇ 5f are dispensed, and among them, the dispensing numbers “A1-01”, “A1-02”, “A1-03”, and “A1-04” are stored in the dispensing numbers of the inspection analysis data 21. This indicates that the petri dishes 5b to 5e indicated by the dispensing numbers “A1-01”, “A1-02”, “A1-03”, “A1-04” are used for the test mobility analysis. ing. The petri dish 5f indicated by the dispensing number “A1-05” is not used in the inspection analysis data 21 shown in FIG.
  • the test chemotaxis result is information indicating the result of the test chemotaxis analysis, and includes columns for the total number of nematodes, positive chemotaxis, negative chemotaxis, neutrality, and chemotaxis index.
  • the total number of nematodes is stored in each of the petri dishes 5b to 5e (test petri dish 51) indicated by the dispensing number.
  • Positive chemotaxis stores the number of nematodes that have attracted the subject's urine.
  • the negative chemotaxis stores the number of nematodes that showed repellent behavior against the subject's urine.
  • Neutral stores the number of nematodes that did not attract or repel the subject's urine.
  • the chemotaxis index an index relating to the nematode chemotaxis calculated by a method described later is stored.
  • the average runtability index is also stored. That is, the average chemotaxis index is an average value of the chemotaxis index for each lot (nematode culture dish 5a; culture unit).
  • positive chemotaxis, negative chemotaxis, neutrality, and the chemotaxis index will be described later.
  • the culture condition ID is an ID for linking to information in the culture condition data 22 described later. That is, the test analysis data 21 is linked to the culture condition data 22 described later by the culture condition ID.
  • the running condition ID is an ID for linking to information in the running condition data 23 described later. That is, the test analysis data 21 is linked to the chemotaxis condition data 23 described later by the chemotaxis condition ID.
  • the quality assurance ID is an ID for linking to information in quality analysis data 24 described later. That is, the inspection analysis data 21 is linked to quality analysis data 24 described later by the quality assurance ID.
  • the quality analysis result is an inspection result by the quality runnability analysis, and is determined based on the runnability index CI in the quality runnability analysis described later in FIG. .
  • the quality analysis result is information stored when the inspection analysis apparatus 4 acquires the quality analysis data 24 described later in FIG. 11 from the quality analysis apparatus 2 via a network (not shown).
  • the lot number, the dispensing number, the culture condition ID, the chemotaxis condition ID, and the quality assurance ID information are input to the administrator or the inspector via the input device 405 (see FIG. 5). It is information input by.
  • the test runnability result and the information on the test result are information input as a result of the runtability analysis process described later.
  • FIG. 9 is a diagram showing a configuration example of culture condition data according to the present embodiment.
  • the culture condition data 22 includes columns for culture condition ID, culture temperature, medium composition, bait, number of days of culture, gene analysis information, and remarks.
  • the culture condition ID is identification information for the culture condition.
  • the culture temperature is a temperature (° C.) in an environment where nematodes are cultured.
  • the culture composition is information indicating the composition of a medium in which nematodes are cultured. Incidentally, description of the medium composition under the culture conditions “1-2” and “1-3” is omitted because it is the same as the culture condition “1-1”.
  • a bait is the information regarding the bait given to a nematode.
  • the number of culture days is the number of days that nematodes are cultured.
  • the date of the gene analysis is stored in the upper stage, and the nematode sequence data obtained as a result of the gene analysis is stored in the lower stage.
  • comments etc. by an administrator or the like are stored. Note that each piece of information in the culture condition data 22 is information input by an administrator or an examiner via the input device 205 (see FIG. 3) or the input device 405 (see FIG. 5).
  • FIG. 10 is a diagram illustrating a configuration example of the running condition data according to the present embodiment.
  • the chemotaxis condition data 23 includes columns of the chemotaxis condition ID, the medium composition, the chemotaxis temperature, the chemotaxis time, the illumination condition, and the remarks.
  • the chemotaxis condition ID is identification information for the chemotaxis condition.
  • the test analysis data 21 and the chemotaxis condition data 23 are linked to each other by the chemotaxis condition ID.
  • the medium composition is information indicating the composition of the medium in the laboratory petri dish 51.
  • the chemotaxis temperature is the temperature (° C.) when the nematode is chemotaxised in the test chemotaxis analysis.
  • the chemotaxis time is the time (minutes) during which the nematode is chemotaxisd in the test chemotaxis analysis.
  • the illumination condition is the brightness of the illumination unit 103 (FIG. 2), and when shooting is performed, which is the brightness when the nematode is running (in the example of FIG. 10, it is turned off), shooting is performed.
  • the brightness at the time of shooting (lit in the example of FIG. 10) is stored.
  • each information of the mobility property data 23 is information input by the administrator or the examiner via the input device 405 (see FIG. 5).
  • FIG. 11 is a diagram illustrating a configuration example of quality analysis data according to the present embodiment.
  • Quality analysis data 24 includes quality assurance ID, chemotaxis temperature, chemotaxis time, lighting conditions, nematode group ID, nematode count, positive chemotaxis, negative chemotaxis, neutrality, chemotaxis index CI, standard substance And each column of the pass line.
  • the quality assurance ID is identification information for the quality runnability analysis result. As described above with reference to FIG. 8, the inspection analysis data 21 and the quality analysis data 24 are linked to each other by the quality assurance ID.
  • the chemotaxis temperature is a temperature (° C.) when the nematode is chemotaxised in the quality chemotaxis analysis.
  • the chemotaxis time is the time (minutes) during which the nematode is chemotaxis in the quality chemotaxis analysis.
  • the illumination condition is the brightness of the illumination unit 103 (FIG. 2) in the quality runnability analysis. In the example of FIG. 11, the illumination unit 103 is always on. However, when the nematode is running, as in FIG. 10, the illumination unit 103 is turned off, and the illumination unit 103 is in shooting. 103 may be lit.
  • the nematode group ID is an ID for linking to information in the nematode data 25 described later. That is, the quality analysis data 24 is linked to the nematode data 25 by the nematode group ID.
  • the total number of nematodes is the total number of nematodes (dispensed in the petri dish 5f) used for quality chemotaxis analysis.
  • Positive chemotaxis is the number of nematodes that have attracted to isoamyl alcohol.
  • Negative chemotaxis is the number of nematodes that showed repellent behavior against isoamyl alcohol.
  • Neutral stores the number of nematodes that did not attract or repel isoamyl alcohol.
  • the chemotaxis index CI an index related to nematode chemotaxis calculated by a predetermined method described later is stored.
  • positive chemotaxis, negative chemotaxis, neutrality, and the chemotaxis index will be described later.
  • the standard substance is a substance used in place of urine in the test chemotaxis analysis, and here, isoamyl alcohol is used.
  • the pass line is a pass line with respect to the result of the quality runnability analysis. In this embodiment, if the runnability index CI is larger than “+0.7” as a result of the quality runnability analysis using isoamyl alcohol as the standard substance, the pass line is passed. It is said. In the example of FIG. 11, the quality assurance IDs “3-1” and “3-2” are acceptable and the quality assurance ID “3-3” is unacceptable.
  • the running condition at the time of quality assurance running is the information of the running condition data 23 linked through the quality assurance ID.
  • the running conditions at the time of quality assurance running with the quality assurance ID “3-1” are as follows.
  • the link source of the quality assurance ID “3-1” is the lot number “A1” in the inspection analysis data 21.
  • the running condition in the lot number “A1” is a condition stored in the running condition ID “2-1” in the running condition data 23. Therefore, the running condition at the time of quality assurance running with the quality assurance ID “3-1” is a condition stored in the running condition ID “2-1” in the running condition data 23.
  • the quality assurance ID, the chemotaxis temperature, the chemotaxis time, the lighting conditions, the nematode group ID, the reference material, and the pass line information are input via the input device 205 (see FIG. 3). Input by the administrator or inspector. Further, the information on the total number of nematodes, positive chemotaxis, negative chemotaxis, neutrality, and chemotaxis index CI is information stored by the chemotaxis analysis process described later.
  • the chemotaxis temperature and the chemotaxis time in the quality chemotaxis analysis and the test chemotaxis analysis are the same. That is, the same chemotaxis condition is used in the quality chemotaxis analysis and the test chemotaxis analysis.
  • a medium composition column may be provided. And the culture medium composition in a quality mobility analysis and a test
  • FIG. 12 is a diagram illustrating a configuration example of nematode data according to the present embodiment.
  • the nematode data 25 is data for managing information on the nematode individual, and the nematode group ID, individual ID, time, ferret diameter, coordinates, travel distance, speed, average ferret diameter, average travel distance, and average speed. It has each column.
  • the nematode group ID is identification information for a group of nematodes used for quality chemotaxis analysis. As described above with reference to FIG. 11, the quality analysis data 24 and the nematode data 25 are linked to each other by the nematode group ID.
  • the individual ID is an ID given to each individual nematode in the nematode group identified by the nematode group ID.
  • Time is the elapsed time (after minutes) when the chemotaxis was performed in the quality chemotaxis analysis.
  • the ferret diameter (mm) is the size shown in FIG. The reason why the ferret diameter is used is that it is difficult to accurately measure the body length (the straight length) of the nematode 701 that is moving at any time. Since the ferret diameter has a size as shown in FIG. 13, the size changes depending on how the nematode grows.
  • the coordinate is the position of the nematode at the corresponding elapsed time.
  • the x coordinate and y coordinate are defined as the positive x coordinate where the center of the petri dish 5f is the origin and the direction of the portion 801 where isoamyl alcohol is dropped (described later in detail) is positive.
  • the y coordinate is defined perpendicular to the defined x coordinate in the plane of the petri dish 5f.
  • the moving distance is the moving distance (mm) of the nematode at the corresponding elapsed time.
  • the speed is the speed (mm / s) of the nematode at the corresponding elapsed time. The speed is calculated by moving distance / elapsed time.
  • the average ferret diameter is the average (mm) of the nematode ferret diameter in the nematode group indicated by the nematode group ID.
  • the average moving distance is the average (mm) of the moving distance of the nematode in the nematode group indicated by the nematode group ID.
  • the average moving speed is the average (mm / s) of the moving speed of the nematode in the nematode group indicated by the nematode group ID.
  • the nematode group ID and the individual ID are information input by the administrator or the examiner via the input device 205 (see FIG. 3). Other information is information calculated and input by the counting unit 211 (see FIG. 3) or the like.
  • FIG. 14 is a flowchart showing the procedure of the runnability analysis process according to this embodiment.
  • This chemotaxis analysis process is a process common to both the test chemotaxis analysis and the quality chemotaxis analysis. Reference is made to FIGS. 2 to 5 as appropriate. Here, the case of quality runnability analysis will be described.
  • the counting unit 11 identifies the position (final position coordinate) of the nematode after electrophoresis (S101). At this time, the counting unit 11 counts the total number of nematodes and inputs the counted result in the column of the total number of nematodes in the inspection analysis data 21 shown in FIG. 8 and the quality analysis data 24 shown in FIG.
  • the counting unit 11 counts nematodes whose final position coordinate x1 is greater than + ⁇ (x1> + ⁇ ) (S102).
  • the number of nematodes counted here is defined as the number N1 of nematodes exhibiting positive chemotaxis (that is, attracting behavior with respect to a standard substance).
  • the counting unit 11 counts the total number of nematodes and inputs the counted result in the column of the positive runnability of the inspection analysis data 21 shown in FIG. 8 and the quality analysis data 24 shown in FIG.
  • the counting unit 11 counts nematodes whose final position coordinate x1 is less than ⁇ (x1 ⁇ ) (S103).
  • the number of nematodes counted here is defined as the number N1 of nematodes exhibiting negative chemotaxis (that is, repellent behavior with respect to a standard substance).
  • the counting unit 11 counts the total number of nematodes, and inputs the counted result in the column of negative mobility of the test analysis data 21 shown in FIG. 8 and the quality analysis data 24 shown in FIG.
  • the counting unit 11 counts nematodes whose final position coordinate x1 is ⁇ or more and + ⁇ or less ( ⁇ ⁇ x1 ⁇ + ⁇ ) (S104).
  • the number of nematodes counted here is defined as the number N1 of nematodes exhibiting neutral chemotaxis.
  • the counting unit 11 counts the total number of nematodes, and inputs the counted result in the neutral column of the inspection analysis data 21 shown in FIG. 8 and the quality analysis data 24 shown in FIG.
  • FIG. 15 is a diagram for describing the chemotaxis determination of a nematode according to the present embodiment.
  • the time of quality runnability analysis shall be demonstrated and the code
  • the nematode chemotaxis determination is performed in the same procedure.
  • Reference numeral 801 denotes a location where isoamyl alcohol + sodium azide is dropped.
  • Reference numeral 802 denotes a portion where sodium azide is dropped. Sodium azide is used as an anesthetic for nematodes.
  • reference numeral 801 is a location where the subject's urine + sodium azide is dropped. As described above, the center of the petri dish 5f is the origin, the direction of the portion 801 where isoamyl alcohol is dropped is defined as a positive x coordinate, and the y coordinate is perpendicular to the defined x coordinate in the plane of the petri dish 5f. Is defined.
  • the nematode is dropped on the center 803 of the petri dish 5f. Since the nematode exhibits an attracting action with respect to isoamyl alcohol, the nematode moves in the positive direction with respect to the x coordinate of FIG.
  • the nematode behaves with respect to the urine. That is, if the urine is from a cancer patient, the nematode moves in the positive direction with respect to the x coordinate in FIG. Conversely, if the urine is from a healthy person, the nematode will repel the urine. That is, if the urine is from a healthy person, the nematode moves in a negative direction with respect to the x coordinate in FIG.
  • a line 811 is a line having a distance of ⁇ in the + direction of the x coordinate from the center 803 of the petri dish 5f and parallel to the y axis.
  • a line 812 is a line that is a distance of ⁇ in the ⁇ direction of the x coordinate from the center 803 of the petri dish 5f and is parallel to the y axis. That is, the number of nematodes in the region 821 where the final position coordinates x1 of the nematodes after the end of the chemotaxis analysis is x1> + ⁇ is the number N1 of nematodes exhibiting positive chemotaxis.
  • the number of nematodes in the region 822 where the final position coordinate x1 is x1 ⁇ is the number N2 of nematodes exhibiting negative chemotaxis.
  • the number of nematodes in the region 823 where the final position coordinate x1 is ⁇ ⁇ x1 ⁇ + ⁇ is the number N3 of nematodes showing neutral chemotaxis.
  • the nematode count is performed, for example, by the following procedure.
  • the counting unit 11 emphasizes the contrast of an image (imaging result) captured by the imaging unit 101 in the quality analysis apparatus 1.
  • A2 The counting unit 11 erases the outline of the petri dish 5f and the culture medium from the image. In this process, after the examiner selects the petri dish 5f in the image or a portion corresponding to the culture medium with a mouse or the like, the image of the selected portion may be erased.
  • the petri dish 5f and the location of the culture medium are the same for all images, so that a location to be erased in the image is set in advance, and the counting unit 11 may erase the set location.
  • the counting unit 11 makes the erased portion and the original background black.
  • the medium may be colored (a color close to black is desirable). In this way, nematodes can be made conspicuous without performing the process (A2).
  • (A3) As a result of the processing of (A2), nematodes are displayed in white granular form on a black background.
  • (A4) The counting unit 11 sets the x-coordinate and the y-coordinate at the vertical and horizontal centers of the image. Further, the counting unit 11 sets + ⁇ and ⁇ lines shown in FIG. 15 in the image. Since the petri dish 5f is photographed in the same size and in the same direction, the counting unit 11 can easily set the x-coordinate, y-coordinate, + ⁇ , and ⁇ lines on the basis of previously input information.
  • the counting unit 11 confirms the final position coordinates of the nematode by counting nematodes displayed in white particles (S101 in FIG.
  • nematode counting methods shown in (A1) to (A5) are merely examples, and other methods may be used.
  • the calculation unit 12 calculates the chemotaxis index CI based on the counting result of the counting unit 11 (S105).
  • the chemotaxis index CI is calculated by the following formula (1).
  • the determination unit 13 determines whether the runnability index CI calculated in step S105 is greater than a predetermined value ⁇ (corresponding to the pass line in FIG. 11) (S106). As a result of step S106, when the runnability index CI is larger than the predetermined value ⁇ (S106 ⁇ Yes), the determination unit 13 determines that there is no problem (S107). That is, the determination unit 13 determines that the reliability of a cancer test using a nematode dispensed from the same lot as the nematode being analyzed is high. As a result of step S106, when the runnability index CI is equal to or less than the predetermined value ⁇ (S106 ⁇ No), the determination unit 13 determines that there is a problem (S108).
  • the determination unit 13 determines that the reliability of the cancer test using a nematode dispensed from the same lot as the nematode being analyzed is low. If it is determined in step S108 that there is a problem, the examiner redoes the cancer test by using a lot different from the nematode lot to be analyzed. By doing in this way, it becomes possible to perform a highly reliable cancer test.
  • the output processing unit 14 describes the determination results of step S107 and step S108 in the inspection result notification 900.
  • the calculation unit 412 of the test analyzer 4 calculates the average value of the chemotaxis index for each lot (nematode culture dish 5a; culture unit), and the calculated average value is shown in FIG. Is stored in the column of the average runnability index in the test analysis data 21 shown in FIG.
  • inspection analyzer 4 determines positive / negative as a result of a cancer test based on the value of an average chemotaxis index.
  • FIG. 16 is a diagram showing an example of a test result notification sheet according to the present embodiment.
  • the inspection result notice 900 is shown to the subject and has an inspection result area 901 and a quality assurance area 902.
  • a cancer test result by nematode (a result of a cancer test by the test analyzer 4) is printed.
  • the cancer test result is printed with reference to the “overall determination” column in FIG.
  • the quality assurance area 902 the result of the quality runnability analysis by the quality analyzer 2 is printed. That is, it is printed that the quality of the nematode used for the cancer test is guaranteed.
  • the quality assurance area 902 the reliability of the subject can be improved.
  • not only the result of the quality runnability analysis but also information on what criteria the quality runtability analysis has been performed may be printed. By doing in this way, the reliability with respect to the cancer test of a subject can further be improved.
  • the cancer test system Z dispenses nematodes from nematodes in the same nematode culture petri dish 5a into a plurality of petri dishes 5b to 5f, of which the nematodes in the petri dishes 5b to 5e are inspected. Used for analysis, nematode in petri dish 5f is used for quality chemotaxis analysis. By doing in this way, the quality of the nematode under the same culture conditions as the nematode (nematode group) used for the cancer test can be analyzed, and the nematode used for the cancer test is suitable for the test. Can be easily confirmed.
  • quality assurance can be further improved by making the test mobility conditions the same in the test mobility analysis and the quality mobility analysis.
  • the work is mainly performed manually, so the results vary depending on the skill level. For example, even if the buffer is wiped after the nematode is dropped on the medium, a difference occurs between the skilled person and the unskilled person. Or if it takes time to dripping sodium azide and nematodes for paralysis of urine and nematodes, the activity of nematodes will be reduced and the test accuracy will be greatly reduced. Thus, even if nematodes of the same condition are used, the inspection accuracy changes depending on the skill level of the work.
  • an actual cancer test is performed using a plurality of petri dishes. That is, multiple drops of sodium azide and nematodes for paralyzing urine and nematodes are performed. As a result, the results of the cancer test indicate what state of the nematode was used, what work was performed, in other words, the state of the nematode used was suitable for the cancer test. Information about what happened is needed.
  • quality assurance is determined based on the attracting behavior or repelling behavior of nematodes. More specifically, quality assurance is determined based on the runnability index CI according to the equation (1). In this way, quantitative quality assurance determination can be performed, and the reliability of quality assurance can be improved.
  • the quality analysis apparatus 1 has the imaging part 101, Based on the image (imaging result information) which the imaging part 101 image
  • the nematode performs an attracting action or a repelling action (what kind of chemotaxis action is known), and what kind of nematode is used It can be confirmed whether it is in a state (activity, etc.). In particular, by using isoamyl alcohol that shows a strong attracting behavior of nematodes, the state (activity, etc.) of the nematodes can be remarkably shown.
  • the quality analysis device 1 and the test analysis device 2 show the test result notification 900 in which the information on quality assurance is described together with the result of the cancer test to the subject, so that the reliability of the cancer test for the subject is confirmed. Can be improved.
  • the quality assurance determination is binarized as “problematic” and “no problem”, but the present invention is not limited to this, and it may be displayed in multiple values in a plurality of ranks.
  • the inspection result notification 900 is printed on paper.
  • the present invention is not limited to this, and may be displayed on a screen of a predetermined PC or the like when the subject logs in. Good.
  • urine as a specimen used for the test run analysis is subjected to a filter process using a sterilization filter or a filter for removing impurities.
  • urine is used as a sample used for the test run analysis.
  • biological materials derived from the subject 2 such as sweat, cells of the subject 2, blood, saliva, exhalation, etc. are used. May be.
  • a wild-type nematode is used, but a genetically modified nematode such as an indicator gene that can measure the calcium concentration in the nerve is expressed in the olfactory nerve of the nematode. It may be used (transgenic nematodes may be used, or knockout nematodes in which a specific gene is disrupted and deleted) may be used.
  • the standard substance (or urine) + sodium azide is dropped on the reference numeral 801 in FIG. 15 and the sodium azide is dropped on the reference numeral 802 in FIG. 15, but this is not restrictive.
  • a standard substance (or urine) + sodium azide may be dropped concentrically around the periphery of the medium.
  • the nematode is dropped on the center 803 of the petri dish 5f in FIG. 15, but the present invention is not limited to this as long as it can measure the chemotaxis behavior of the nematode.
  • nematodes may be sprayed on the medium in the petri dish 5f to measure how much nematodes gather around the standard substance (or urine) + sodium azide.
  • each of the above-described configurations and functions, each of the units 11 to 14, 211 to 214, 411 to 414, the storage devices 202 and 402, etc. are implemented by hardware by designing a part or all of them, for example, with an integrated circuit It may be realized.
  • the above-described configurations, functions, and the like may be realized by software by interpreting and executing programs that realize the respective functions by the processors such as the CPUs 203 and 403. .
  • information such as programs, tables, and files for realizing each function is stored in the HDD, as well as a memory 201, 401, a recording device such as an SSD, or an IC (Integrated Circuit).
  • control lines and information lines are those that are considered necessary for explanation, and not all control lines and information lines are necessarily shown on the product. In practice, it can be considered that almost all configurations are connected to each other.

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Abstract

Selon l'invention, afin de réaliser l'analyse de la qualité d'un ver nématode dans un test de cancer utilisant le ver nématode de manière simple, un système de test de cancer est caractérisé en ce qu'il comprend : un dispositif d'analyse de qualité (1), dans lequel une substance standard est déposée sur un emplacement spécifique dans une boîte de pétri spécifique (5f) parmi de multiples boîtes de pétri (5b à 5f) ayant chacune le ver nématode préparé à l'intérieur de ces dernières à partir d'une boîte de pétri unique (5a), et dans lequel le comportement de migration du ver nématode en réponse à la substance standard est imagé à chaque temps écoulé prédéterminé ; un dispositif d'évaluation de qualité (2), dans lequel le comportement d'attraction et le comportement d'évitement du ver nématode en réponse à la substance standard est évalué sur la base d'informations envoyées par le dispositif d'analyse de qualité (1), puis des informations sur la qualité du ver nématode sont délivrées en sortie sur la base du résultat du test ; et un dispositif d'analyse de test 3 et un dispositif d'évaluation de test 4, dans chacun desquels le test sur le cancer est effectué à l'aide de boîtes de pétri (5b à 5e) qui sont différentes de la boîte de pétri (5f) pour une utilisation en analyse de qualité/migration.
PCT/JP2015/081611 2015-11-10 2015-11-10 Système de test de cancer et procédé d'évaluation de test de cancer Ceased WO2017081750A1 (fr)

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WO2022219764A1 (fr) * 2021-04-15 2022-10-20 陶徳堂有限公司 Procédé de détermination, dispositif de mesure et système de mesure
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JP7442785B2 (ja) 2019-12-06 2024-03-05 株式会社Ｈｉｒｏｔｓｕバイオサイエンス 走性解析方法、がん評価方法、走性解析システム及びプログラム
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JP2021135265A (ja) * 2020-02-28 2021-09-13 陶徳堂有限公司 判定方法、測定装置、及び測定システム
JPWO2022107736A1 (fr) * 2020-11-17 2022-05-27
WO2022107736A1 (fr) * 2020-11-17 2022-05-27 株式会社Ｈｉｒｏｔｓｕバイオサイエンス Méthode de test d'échantillons sanguins utilisant le comportement de taxie d'un nématode
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WO2022219764A1 (fr) * 2021-04-15 2022-10-20 陶徳堂有限公司 Procédé de détermination, dispositif de mesure et système de mesure
EP4392775A4 (fr) * 2021-08-24 2025-08-06 Oncotect Inc Dépistage du cancer chez des animaux de compagnie

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